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Recombinase Polymerase Amplification Combined with a Lateral Flow Strip for the Detection of Plasmodium knowlesi
Meng-Yee Lai
Meng-Yee Lai
Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia;
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Choo-Huck Ooi
Choo-Huck Ooi
Sarawak State Health Department, Kuching, Sarawak, Malaysia
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Yee-Ling Lau
Yee-Ling Lau
Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia;
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The aim of this study was to develop a recombinase polymerase amplification (RPA) combined with a lateral flow (LF) strip method for specific diagnosis of Plasmodium knowlesi. With incubation at 37°C, the 18S rRNA gene of P. knowlesi was successfully amplified within 12 minutes. By adding a specifically designed probe to the reaction solution, the amplified RPA product can be visualized on a LF strip. The RPA assay exhibited high sensitivity with limits of detection down to 10 parasites/μL of P. knowlesi. Nonetheless, it was demonstrated that all P. knowlesi (N = 41) and other Plasmodium sp. (N = 25) were positive while negative samples (N = 8) were negative. Therefore, a combination of RPA and LF strip detection is a highly promising approach with the potential to be suitable for use in resource-limited settings.
Author Notes
Financial support: This study was supported by University Malaya PPP Research Grant (PG066-2016A).
Authors’ addresses: Meng-Yee Lai and Yee-Ling Lau, Department of Parasitology, Faculty of Medicine, University of Malaya, Kuala Lumpur, Malaysia, E-mails: mengylai11@yahoo.com and lauyeeling@um.edu.my. Choo-Huck Ooi, Sarawak State Health Department, Kuching, Sarawak, Malaysia, E-mail: ooi.choo.huck@gmail.com.
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
World Health Organization, 2016. The World Malaria Report 2016. Geneva, Switzerland: World Health Organization.
-
2.
Oriero EC, Geertruyden JV, Nwakanma DC, D’Alessandro U, Jacobs J, 2015. Novel techniques and future directions in molecular diagnosis of malaria in resource-limited settings. Expert Rev Mol Diagn 15: 1419–1426.
-
3.
Singh B, 2016. Plasmodium knowlesi: an update. Microbiol Aust 37: 39–42.
-
4.
Richards J, Mueller I, 2017. Identifying the risks for human transmission of Plasmodium knowlesi. Lancet Plenet Health 1: e83–e85.
-
5.
Willmann M, Ahmed A, Siner A, Wong IT, Lu CW, Singh B, Krishna S, Cox-Singh J, 2012. Laboratory markers of disease severity in Plasmodium knowlesi infection: a case control study. Malar J 11: 363.
-
6.
Piepenburg O, Williams CH, Stemple DL, Armes NA, 2006. DNA detection using recombination protein. PLoS Biol 4: e204.
-
7.
Kersting S, Rausch V, Bier FF, von Nickisch-Rosenegk M, 2014. Rapid detection of Plasmodium falciparum with isothermal recombinase polymerase amplification and lateral flow analysis. Malar J 13: 99.
-
8.
Liu J, Mazumdar D, Lu Y, 2006. A simple and sensitive “dipstick” test in serum based on lateral flow separation of aptamer-linked nanostructures. Angew Chem Int Ed Engl 5: 7955–7959.
-
9.
Posthuma-Trumpie GA, Korf J, Amerongen A, 2009. Lateral flow (immuno) assay: its strengths, weaknesses, opportunities and threats. A literature survey. Anal Bioanal Chem 393: 569–582.
-
10.
Ohrt C, O’Meara W, Remich S, McEvoy P, Ogutu B, Mtalib R, Odera J, 2008. Pilot assessment of the sensitivity of the malaria thin film. Malar J 7: 22.
-
11.
Moody A, 2002. Rapid diagnostic tests for malaria parasites. Clin Microbiol Rev 5: 66–78.
-
12.
Iseki H, Kawai S, Takahashi N, Hirai M, Tanabe K, Yokoyama N, Igarashi I, 2010. Evaluation of a loop-mediated isothermal amplification method as a tool for diagnosis of infection by the zoonotic simian malaria parasite Plasmodium knowlesi. J Clin Microbiol 48: 2509–2514.
-
13.
Lau YL, Lai MY, Anthony CA, Chang PY, Palaeya V, Fong MY, Mahmud R, 2015. Comparison of three molecular methods for the detection and speciation of five human Plasmodium species. Am J Trop Med Hyg 92: 28–33.
-
14.
Divis PCS, Shokoples SE, Singh B, Yanow SK, 2010. A TaqMan real-time PCR assay for the detection and quantitation of Plasmodium knowlesi. Malar J 9: 344.
-
15.
Al-Soud WA, Radstrom P, 2001. Purification and characterization of PCR inhibitory components in blood cells. J Clin Microbiol 39: 485–493.
-
16.
Garcia ME, Blanco JL, Caballero J, Gargallo-Viola D, 2002. Anticoagulants interfere with PCR used to diagnose invasive aspergillosis. J Clin Microbiol 40: 1567–1568.
-
17.
Weyant RS, Edmonds P, Swaminathan B, 1990. Effect of ionic and nonionic detergents on the Taq polymerase. Biotechniques 9: 308–309.
-
18.
Li Y, Kumar N, Gopalakrishnan A, Ginocchio C, Manji R, Bythrow M, Lemieux B, Kong H, 2013. Detection and species identification of malaria parasites by isothermal tHDA amplification directly from human blood without sample preparation. J Mol Diagn 15: 634–641.
| Past two years | Past Year | Past 30 Days | |
|---|---|---|---|
| Abstract Views | 547 | 459 | 53 |
| Full Text Views | 865 | 21 | 2 |
| PDF Downloads | 337 | 17 | 2 |